Oral compositions for inhibiting lipid accumulation, oral compositions for inhibiting obesity, and oral compositions for anti-inflammation, as well as functional foods, supplements, and pharmaceutical compositions

Abstract

To provide an oral composition having an excellent lipid accumulation inhibiting effect.SOLUTION: A lipid accumulation inhibiting oral composition contains a Litsea cubeba leaf extract and / or its dried matter. The oral composition has an excellent lipid accumulation inhibiting effect and can be desirably used for functional foods, supplements, pharmaceutical compositions, etc.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to an oral composition for suppressing lipid accumulation, an oral composition for suppressing obesity, and an oral composition for anti-inflammation, and their application products.

Background Art

[0002] Obesity is generally a state in which not only the weight is higher than the normal state but also excessive lipids are accumulated in adipose tissue. The accumulation of lipids occurs when the balance between the intake energy obtained from diet or the like and the consumption energy consumed by basal metabolism, physical activity, or the like is not maintained. That is, when basal metabolism or the like decreases, the intake energy becomes surplus as a whole body, and that portion becomes fat (subcutaneous fat or visceral fat) and accumulates. Obesity is caused by lifestyle habits such as overeating and lack of exercise, and the extra energy is accumulated as body fat because the intake energy is more than the consumption energy. Obesity is a cause of metabolic syndrome and lifestyle-related diseases, and is also associated with chronic inflammatory diseases such as diabetes, arteriosclerosis, and cancer. In order to prevent such diseases, it is necessary to eliminate obesity.

[0003] So far, various oral compositions for suppressing obesity have been reported. For example, an obesity improver used for obesity caused by lipid accumulation, containing Stephaniae Tetrandrae-Radix Astragali Decoction extract (Patent Document 1), a dietary composition for improving obesity obtained by mixing an extract obtained from the leguminous plant Phaseolus calcaratus Roxb. and an extract obtained from the moraceous plant Morus alba L. (Patent Document 2), an obesity prevention and improvement agent containing an aqueous component of defatted meal of fruits and / or seeds of Camellia japonica belonging to the genus Camellia of the family Theaceae as an active ingredient (Patent Document 3), etc. have been reported.

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Patent Document 2

Patent Document 3

Summary of the Invention

Problems to be Solved by the Invention

[0005] By the way, the suppression of obesity by the intake of an oral composition is based on the action of decomposing lipids already accumulated in adipose tissue (lipolytic action) by the components contained in the oral composition, or the action of suppressing the new accumulation of lipids in adipose tissue (lipid accumulation inhibitory action). Here, the lipid accumulation inhibitory action suppresses the accumulation of lipids in the body, and thus contributes to the prevention of obesity. Therefore, from the viewpoint of preventing obesity, the lipid accumulation inhibitory action is particularly important rather than the lipolytic action. Therefore, components having an excellent lipid accumulation inhibitory action have been searched for.

[0006] Under such circumstances, an object of the present invention is to provide an oral composition useful for suppressing lipid accumulation and its application products.

Means for Solving the Problems

[0007] As a result of intensive studies to solve the above problems, the present inventors have found that the extract and dried product of Momordica charantia leaves have a lipid accumulation inhibitory action, and have reached the present invention.

[0008] That is, the present invention relates to the following inventions. <1> An oral composition for suppressing lipid accumulation containing an extract and / or dried product of Momordica charantia leaves. <2> The oral composition according to <1>, wherein the extract of Momordica charantia leaves is a solvent extract of Momordica charantia leaves. <3> The oral composition according to <2>, wherein the solvent extract of Momordica charantia leaves is a solvent extract using a mixed solvent of water and ethanol of Momordica charantia leaves as an extraction solvent. <4> The oral composition according to <2>, wherein the solvent extract of Momordica charantia leaves is a hot water extract of Momordica charantia leaves. <5> An oral composition for suppressing obesity containing an extract and / or a dried product of blue mold leaves. <6> An oral composition for anti-inflammation containing an extract and / or a dried product of blue mold leaves. <7> A functional food containing the oral composition according to any one of <1> to <6>. <8> A supplement containing the oral composition according to any one of <1> to <6>. <9> A pharmaceutical composition containing the oral composition according to any one of <1> to <6>.

[0009] <1a> A method for producing an oral composition for suppressing lipid accumulation, which has an extraction step of obtaining an extract of blue mold leaves by bringing blue mold leaves into contact with an extraction solvent of a mixed solvent of water and ethanol. <2a> A method for producing an oral composition for suppressing lipid accumulation, which has an extraction step of obtaining an extract of blue mold leaves by bringing blue mold leaves into contact with hot water.

Effect of the Invention

[0010] According to the present invention, an oral composition useful for suppressing lipid accumulation and its application products are provided.

Brief Description of the Drawings

[0011]

Figure 1

Figure 2

Figure 3

Figure 4

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Figure 6

Figure 7

Mode for Carrying Out the Invention

[0012] Hereinafter, the present invention will be described in detail with reference to examples and the like. However, the present invention is not limited to the following examples and the like, and can be arbitrarily modified and implemented without departing from the gist of the present invention. In this specification, "~" is used as an expression including the numerical values or physical quantities before and after it. Also, in this specification, the expression "A and / or B" includes "only A", "only B", and "both A and B".

[0013] <1. Composition for Suppressing Lipid Accumulation> The present invention relates to an oral composition for suppressing lipid accumulation containing an Aomoji leaf extract and / or a dried product (hereinafter referred to as "the oral composition for suppressing lipid accumulation of the present invention"). In this specification, the Aomoji leaf extract and / or the dried product may be collectively referred to as "processed Aomoji leaves".

[0014] As shown in the examples described later, the processed Aomoji leaves have an effect of suppressing lipid accumulation. Therefore, the processed Aomoji leaves can be used as an active ingredient of an oral composition for suppressing lipid accumulation. Note that there has been no report on the lipid accumulation suppressing effect of the processed Aomoji leaves so far.

[0015] As used herein, the "lipid accumulation inhibitory effect" means an effect of suppressing lipid accumulation in adipocytes. Note that the lipid accumulation inhibitory effect can be clearly distinguished from the effect of decomposing lipids already accumulated in adipocytes (lipid decomposition effect). However, it is not intended to deny that the processed aomorogi leaf according to the present invention has a lipid decomposition effect, and the oral composition for suppressing lipid accumulation of the present invention may have a lipid decomposition effect together with the lipid accumulation inhibitory effect.

[0016] The lipid accumulation inhibitory effect can be determined by evaluating the amount of lipid accumulation in adipocytes and the weight of adipose tissue around organs in the living body of a mouse. Specifically, it can be evaluated by the evaluation methods <3-1. Evaluation of lipid accumulation in cells> and <3-2. Evaluation of lipid accumulation in the living body of a mouse> described in the examples below.

[0017] Since the processed aomorogi leaf according to the present invention has a lipid accumulation inhibitory effect, an oral composition containing the processed aomorogi leaf is useful for suppressing obesity. Therefore, a composition containing the processed aomorogi leaf can be used as an oral composition for suppressing obesity.

[0018] Here, as used herein, "obesity suppression" is not particularly limited as long as it has the meaning used in the art, and includes eliminating or alleviating a state in which excessive lipids are accumulated in the body (obesity) (improvement of obesity) and suppressing the progression of obesity (prevention of obesity). Therefore, the oral composition for suppressing obesity of the present invention is useful not only for suppressing the increase in body weight due to lipid accumulation and alleviating the increase in body weight, but also for maintaining a normal body weight.

[0019] In addition to the lipid accumulation inhibitory effect described above, the oral composition according to the present invention also has an effect of suppressing sugar absorption in small intestinal epithelial cells and an effect of promoting sugar uptake in liver cells. Therefore, the oral composition according to the present invention can be expected to have an effect of suppressing obesity from both lipid accumulation inhibition and sugar absorption inhibition. Here, in this specification, the "sugar absorption action in intestinal epithelial cells" means the action of absorbing sugar through the sugar transport protein (SGLT1) present in intestinal epithelial cells. The "sugar uptake action in liver cells" means the action of taking up sugar in the liver by the action of insulin after a meal.

[0020] The oral composition for suppressing lipid accumulation (and the oral composition for suppressing obesity) of the present invention exhibits a lipid accumulation suppressing action when orally ingested, and thus may be used as a pharmaceutical composition for suppressing lipid accumulation (for suppressing obesity), or may be used as a functional food or supplement for suppressing lipid accumulation (for suppressing obesity). The oral composition for suppressing lipid accumulation of the present invention is useful for at least one of prevention, treatment, and improvement of symptoms of diseases and symptoms (neurodegenerative diseases, cardiovascular diseases, obesity, etc.) caused by lipid accumulation in the body. That is, "prevention" includes suppression and delay of onset of the disease or symptom. Also, "treatment" includes improvement and remission of the pathological condition of the disease or symptom, and suppression of the progression of the disease or symptom.

[0021] Hereinafter, the composition for suppressing lipid accumulation of the present invention will be described in more detail.

[0022] Litsea cubeba used as a raw material of the composition for suppressing lipid accumulation of the present invention is a plant of the genus Litsea in the Lauraceae family, and the part of Litsea cubeba used in the present invention is the leaf part. The Litsea cubeba leaves used as a raw material can be used as a raw material at any time during the cultivation period as long as the effects of the present invention are not impaired. The Litsea cubeba leaves may be natural products or may be artificially cultivated. The composition for suppressing lipid accumulation of the present invention only needs to contain Litsea cubeba leaves (the leaf part of Litsea cubeba), and may contain other parts as long as the effects of the present invention are not impaired.

[0023] The processed Litsea cubeba leaves are a general term for the Litsea cubeba leaf extract and / or dried product as described above. In the present invention, the "extract" is a general concept encompassing forms in which the content of active ingredients has been increased by subjecting the target raw material plant, or a dried and minced form thereof as necessary, to pressing, solvent extraction, or the like. Specifically, it includes any of an extract obtained using a momiji leaf as an extraction raw material, a diluted or concentrated solution of the extract, or a crude or purified product thereof. Note that a dried product obtained by drying the extract also falls within the scope of the extract.

[0024] In the present invention, the "dried product" is a dried raw material plant. Specifically, it is a momiji leaf dried by a known drying method such as sun drying, heat drying, or freeze drying. The drying conditions are not particularly limited as long as the effects of the present invention are not impaired. From the perspective of usability, the dried product is usually pulverized into a powder and used as a dried pulverized product. The pulverization method is not particularly limited, and a conventionally known pulverizer may be used. The particle size of the powder is appropriately determined according to its usage mode.

[0025] The momiji leaf extract and the momiji leaf dried product may each be used alone, or they may be mixed and used in any ratio.

[0026] Among the processed momiji leaf products according to the present invention, a solvent extract is preferred. The solvent extract of momiji leaf can be obtained by extracting the leaf part of momiji according to a conventional method. The extraction method is not particularly limited as long as the effects of the present invention are not impaired, and an appropriate method can be arbitrarily selected and used according to the purpose. For example, a method in which momiji leaf as it is or a processed product such as its crushed product, dried product, or pulverized product is brought into contact with an extraction solvent to extract the components contained in the momiji leaf is preferred.

[0027] Note that the extraction solvent may contain additives within a range that does not impair the effects of the present invention. Examples include a pH adjuster, a viscosity adjuster, and the like.

[0028] A preferred example of the solvent extract of perilla leaves is a solvent extract obtained by using a mixed solvent of water and ethanol as the extraction solvent for perilla leaves. The solvent extract can be safely orally ingested and has an excellent lipid accumulation inhibitory effect. The ratio of water to ethanol in the mixed solvent is arbitrary as long as the effects of the present invention are achieved. For example, when the mixed solvent of water and ethanol is 100% by volume, the ratio of ethanol is 0.1 to 95% by volume.

[0029] A preferred example of the solvent extract of perilla leaves is an aqueous extract of perilla leaves. The water used for extraction includes distilled water, pure water, and sterilized water. For example, tap water or water containing impurities may be used, but water that does not contain components that interfere with the extraction of perilla leaves is preferred. As the water as the extraction solvent, cold water, normal temperature water, warm water, hot water, etc. can be used as long as the effects of the present invention are not impaired, but hot water is preferred from the viewpoint of efficiently extracting components having an excellent lipid accumulation inhibitory composition. The hot water extract of perilla leaves can be safely orally ingested and has an excellent lipid accumulation inhibitory composition.

[0030] The method for extracting the solvent extract from perilla leaves may be any method that can extract the components that exhibit the lipid accumulation inhibitory effect contained in perilla leaves. To illustrate a preferred method for producing a perilla leaf extract, there is a method of bringing perilla leaves into contact with an extraction solvent and stirring for a certain period of time until the active ingredient is sufficiently extracted from the perilla leaves into the extraction solvent.

[0031] The extraction time is appropriately selected in consideration of the type of extraction solvent and the ratio of perilla leaves to the extraction solvent. However, as long as the effects of the present invention are not impaired and the active ingredient can be extracted by the extraction solvent, it is not particularly limited. Also, the obtained extract may be used as it is, or may be subjected to treatments such as dilution, concentration, drying, and purification according to conventional methods. Further, the solvent may be removed by concentration under reduced pressure or freeze-drying as necessary.

[0032] <2. Application of Oral Composition> The oral composition for suppressing lipid accumulation of the present invention may use the processed product of Momordica charantia leaves as it is in the present invention. Usually, other components that do not impair the effects of the present invention are combined, and it is used as an oral composition containing the processed product of Momordica charantia leaves (hereinafter, may be described as "the oral composition of the present invention"). As other components, any components that can be safely ingested by humans may be used, and they are appropriately determined in consideration of the usage forms described later (for example, supplements and functional foods described later). The blending ratio of other components can be appropriately selected and determined according to the purpose as long as the effects of the present invention are not impaired.

[0033] As described above, the oral composition containing the processed product of Momordica charantia leaves according to the present invention exhibits excellent effects on the prevention and improvement of obesity due to its lipid accumulation inhibitory action and sugar absorption inhibitory action. Therefore, the oral composition of the present invention can be used as an oral composition for suppressing obesity, and can also be individually used as an oral composition for suppressing lipid accumulation or for suppressing sugar absorption.

[0034] Furthermore, as shown in the examples described later, the oral composition containing the processed product of Momordica charantia leaves according to the present invention has an anti-inflammatory action, and thus can be used as an "oral composition for anti-inflammation".

[0035] In this specification, the "anti-inflammatory action" means an action of suppressing inflammation in the body. It is known that inflammation, when it becomes chronic, causes damage to living tissues and a decrease in functions such as immunity, and is the cause of diseases such as arteriosclerosis and diabetes caused by these. Therefore, suppressing inflammation contributes to the prevention and treatment of diseases such as arteriosclerosis and diabetes.

[0036] The anti-inflammatory action can be judged by evaluating the inhibitory action of the production of nitric oxide (NO) in macrophage cells. Specifically, it can be evaluated by the evaluation method <3-3. Evaluation of anti-inflammatory action> of the examples described later.

[0037] As described in the examples below, the oral composition containing the processed ao-moji leaves has an excellent anti-inflammatory effect and can thus be used as an oral composition for anti-inflammatory purposes.

[0038] The oral composition of the present invention has an excellent effect on suppressing the production of nitric oxide (NO) due to the contained processed ao-moji leaves. Therefore, as described above, in addition to being used as an oral composition for anti-inflammatory purposes, it can also be individually used as an "oral composition for suppressing nitric oxide production".

[0039] Since the oral composition of the present invention has an excellent lipid accumulation inhibitory effect and anti-inflammatory effect, it can be used in supplements, functional foods, pharmaceutical compositions, etc. for lipid accumulation inhibition (obesity inhibition) and / or anti-inflammatory purposes.

[0040] When the oral composition of the present invention is used for lipid accumulation inhibition, the amount of the processed ao-moji leaves contained is appropriately determined within a range that allows the necessary amount to be ingested, taking into consideration the type and degree of symptoms involved in lipid accumulation, the form of the composition of the present invention, and the method of use.

[0041] When the oral composition of the present invention is used for anti-inflammatory purposes, the amount of the processed ao-moji leaves contained is appropriately determined within a range that allows the necessary amount to be ingested, taking into consideration the type and degree of symptoms involved in inflammation, the form of the composition of the present invention, and the method of use.

[0042] The intake amount of the oral composition of the present invention is not particularly limited as long as it can exhibit the effects of the present invention, and is appropriately determined according to the form of the oral composition of the present invention, the gender, body weight, health condition, etc. of the subject to be ingested. The intake amount may be taken once or divided into several times a day. Also, in order to obtain the effects or functions of the oral composition of the present invention, it is preferable to continuously ingest the oral composition of the present invention.

[0043] Hereinafter, supplements, functional foods, food additives, and pharmaceutical compositions, which are preferred forms of the oral composition of the present invention, will be described, but the uses of the oral composition of the present invention are not limited thereto.

[0044] The oral composition (processed ao-modi leaves) of the present invention can be mixed with various foods and beverages to form a functional food so that it can be easily taken orally on a daily basis, and it is also easy to take it over a long period of time. The "functional food" referred to here means foods and / or beverages ingested for the purpose of maintaining health in addition to general foods, and is a concept including health functional foods (foods for specified health use, nutritional functional foods or foods with function claims), health foods, dietary supplements, nutritional insurance foods, etc. Among these, foods for specified health use, nutritional functional foods, and foods with function claims, which are health functional foods, are preferred embodiments of functional foods. When commercialized as a functional food, various additives used in foods, specifically, coloring agents, preservatives, thickening stabilizers, antioxidants, bleaching agents, antibacterial and antifungal agents, sour agents, seasonings, emulsifiers, fortifiers, manufacturing agents, flavors, etc. may be added.

[0045] The foods and beverages that are the targets of functional foods are not particularly limited. For example, as foods, there are mentioned foods such as sausages, hams, processed fishery products, jellies, candies, chewing gums, etc. Also, as beverages, there are mentioned various teas, green juice beverages, soft drinks, alcoholic beverages, nutritional drinks, etc.

[0046] The ratio of the processed ao-modi leaves to be blended into the functional food is arbitrary, but the blending ratio is selected within a range that can contribute to the lipid accumulation inhibitory action and anti-inflammatory action.

[0047] In addition, the oral composition (processed ao-modi leaves) of the present invention can also be used in the form of a supplement. The form of the supplement is not particularly limited and may be any form such as tablets, powders, granules, capsules, sugar-coated tablets, films, troches, chewable tablets, solutions, emulsions, suspensions, etc. In addition to the composition of the present invention, the supplement of the present invention may contain any components commonly used as supplements. Such components include, for example, amino acids, peptides; vitamins such as vitamin E, vitamin C, vitamin A, vitamin B, folic acid; minerals; saccharides; inorganic salts; citric acid or its salts; tea extracts; fats and oils; nourishing and strengthening components such as propolis, royal jelly, taurine; crude drug extracts such as ginger extract, ginseng extract; herbs; collagen, etc.

[0048] The proportion of the processed aomodori leaf to be blended into the supplement is arbitrary, but the blending proportion is selected within the range that can contribute to the lipid accumulation inhibitory action and anti-inflammatory action.

[0049] When the oral composition of the present invention is used as a functional food, the actions or effects brought about by the processed aomodori leaf according to the present invention may be indicated in the product. For example, it includes the display on the main body, packaging, container, stored items such as packages, or advertisements of the product according to the present invention.

[0050] Further, the oral composition (processed aomodori leaf) of the present invention can be used as a food additive by itself or by adding other components thereto. Other components are not particularly limited as long as they can be used as food and beverage additives. The beverages and foods to which the food additive is added are also arbitrary and not particularly limited.

[0051] The proportion of the oral composition of the present invention to be blended into the food additive is arbitrary, but the blending proportion is selected within the range that can contribute to the lipid accumulation inhibitory action and anti-inflammatory action.

[0052] The oral composition of the present invention can also be applied to animals other than humans. Therefore, it can also be made into an oral composition for animals such as pet food.

[0053] (Pharmaceutical composition) Since the oral composition of the present invention has an action of suppressing lipid accumulation and an anti-inflammatory action, the oral composition of the present invention can be formulated into an oral composition for pharmaceutical use (hereinafter sometimes referred to as "the pharmaceutical composition of the present invention") by mixing an effective amount thereof with a pharmaceutically acceptable base material.

[0054] As used herein, the term "pharmaceutical composition" means a drug useful for at least one of prevention, treatment, and improvement of symptoms of a target disease. In addition, as used herein, the "pharmaceutical composition" includes not only pharmaceuticals but also quasi-drugs.

[0055] The pharmaceutical composition of the present invention may contain other drugs or any pharmacologically acceptable components as long as its efficacy is not impaired in addition to the processed aomori leaf product.

[0056] The pharmaceutical composition of the present invention is useful for at least one of prevention, treatment, and improvement of symptoms of diseases involving lipid accumulation and inflammation. That is, "prevention" includes suppression and delay of onset of the disease or symptom. In addition, "treatment" includes improvement and remission of the pathological condition of the disease or symptom, and suppression of progression of the disease or symptom.

[0057] There is no limitation on the diseases or symptoms targeted by the pharmaceutical composition of the present invention, but specific examples include, for example, dyslipidemia, hypertension, hyperlipidemia, liver diseases, atherosclerotic diseases (stroke and myocardial infarction), neurodegenerative diseases (Alzheimer's disease and Parkinson's disease), cancer, diabetes, inflammatory diseases, rheumatoid arthritis, and the like.

[0058] The pharmaceutical composition of the present invention only needs to contain the processed aomori leaf product, and may be in any form of drug. Examples of the form of the pharmaceutical composition of the present invention include dosage forms such as tablets, pills, granules, powders, liquids, suspensions, syrups, or capsules. The form of the pharmaceutical composition of the present invention may be a solid or a liquid, or a combination of a solid and a liquid, and is not particularly limited. A preferred embodiment of the pharmaceutical composition of the present invention is an oral preparation.

[0059] The pharmaceutical composition of the present invention can also be applied to animals other than humans. Therefore, it can also be used as a pharmaceutical composition for animals.

[0060] The intake amount of the pharmaceutical composition of the present invention is not particularly limited as long as it can exert the effects of the present invention, and is appropriately determined according to the gender, body weight, health status, etc. of the subject to be ingested. The intake amount may be taken once or divided into several times a day. Also, from the viewpoints of preventing, treating, and improving symptoms of the target disease, it is preferable to continuously ingest the pharmaceutical composition of the present invention.

Examples

[0061] The present invention will be described more specifically with reference to the following examples, but the present invention is not limited thereto. In addition, “%” shown in the mixed solvents of the examples indicates “volume %” unless otherwise specified. Also, when the extraction solvent is a mixed solvent, the balance water is omitted from the description. For example, when described as “50% ethanol”, it means a mixed solvent of 50 volume % ethanol and 50 volume % water.

[0062] <1. Preparation of dried aomoji leaf> The collected aomoji leaves were washed with tap water and then blanched for 5 minutes using a steam convection oven (humidity 100%, temperature 100°C). Next, drying was performed for 20 hours using a dryer (dry bulb 65°C, wet bulb 35°C). The dried aomoji leaves were pulverized so as to pass through a 0.4 mm mesh to obtain a dried and pulverized aomoji leaf product.

[0063] <2. Preparation of aomoji leaf extract> (Experimental Example 1) Extraction 1 from dried aomoji leaf with 50% ethanol 4 g of dried and ground aomorji leaves were added with 400 mL of 50% ethanol solution, and after stirring extraction was carried out at room temperature for 3 hours, an extract was obtained by a centrifuge (10,000×g, 30 minutes, 4 °C). Further, 400 mL of 50% ethanol solution was added again to the extraction residue, and after stirring extraction was carried out at room temperature for 3 hours, an extract was obtained by a centrifuge (10,000×g, 30 minutes, 4 °C). The obtained extracts were combined and then dried using a rotary evaporator, a centrifugal concentrator and a freeze dryer to obtain the aomorji leaf extract of Experimental Example 1 in the form of a dry powder.

[0064] (Experimental Example 1a) Extraction of aomorji leaves with 50% ethanol from dried aomorji leaves 2 The same dried and ground aomorji leaves as in Experimental Example 1 were used. 0.5 g of the dried and ground aomorji leaves were immersed in 10 mL of 50% ethanol solution, and after shaking extraction (1,500 rpm) was carried out for 1 hour, the aomorji leaf extract of Experimental Example 1a was obtained by a centrifuge (10,000×g, 30 minutes, 4 °C).

[0065] (Experimental Example 2) Extraction of aomorji leaves with hot water from dried aomorji leaves The same dried and ground aomorji leaves as in Experimental Example 1 were used. 0.5 g of the dried and ground aomorji leaves were immersed in 35 mL of pure water heated to 85 °C, allowed to stand for 5 minutes, and after immersion extraction (85 °C) was carried out, the aomorji leaf extract of Experimental Example 2 was obtained by filtration using qualitative filter paper No. 2.

[0066] <3-1. Evaluation of lipid accumulation in cells> The mouse preadipocyte cell line 3T3-L1 cells were obtained from the Human Science Promotion Foundation Research Resource Bank (cell number JCRB9014) and cultured under the conditions of 37 °C and 5% CO2 using a culture medium (D-biotin 3.2 μM, calcium pantothenate 16 μM, low glucose Dulbecco's Modified Eagle Medium (D-MEM) containing 10% fetal bovine serum). The cells were 2.0×10 4 cells / cm 2They were seeded in a 24-well plate at a density of , and after 2 days, the medium was replaced with a differentiation medium (high-glucose D-MEM containing 3.2 μM D-biotin, 16 μM calcium pantothenate, 0.5 mM 3-isobutyl-1-methylxanthine (IBMX), 0.25 μM dexamethasone (DEX), 10 μM insulin, and 10% fetal bovine serum (FBS)) to induce differentiation. Two days after the induction of differentiation, the medium was replaced with a maturation medium (high-glucose D-MEM containing 3.2 μM D-biotin, 16 μM calcium pantothenate, 10 μM insulin, and 10% FBS). The medium was changed every other day during the maturation period. The extract of Aomodi leaves (Experimental Example 1 or Experimental Example 2) was mixed into the medium at final concentrations of 0, 25, 50, 100, and 200 μg / mL. The culture was terminated 8 days after the start of maturation.

[0067] The amount of intracellular lipid accumulation was measured by Oil Red O staining. Oil Red O was dissolved in 2-propanol to a concentration of 0.5 mg / mL, and water was mixed at a ratio of 3:2 to prepare an Oil Red O staining solution. After the culture was terminated, 4% paraformaldehyde phosphate buffer was added and left standing at room temperature for 1 hour to fix the cells. After removing the 4% paraformaldehyde phosphate buffer, the Oil Red O staining solution was added and left standing at room temperature for 15 minutes. After removing the Oil Red O staining solution and washing the cells, the dye was eluted with 2-propanol. 100 μL of the dye eluate was transferred to a 96-well plate, and the absorbance at 540 nm was measured with a microplate reader (Molecular Devices, LLC, CA, USA) to evaluate the amount of intracellular lipid accumulation.

[0068] Taking the lipid accumulation amount in the group without the extract of Aomodi leaves (50% ethanol extraction or hot water extraction) (0 μg / mL) as 1, the relative value of the lipid accumulation amount with the addition of the extract of Aomodi leaves (50% ethanol extraction or hot water extraction) was calculated. The results are shown in FIGS. 1(a) and 1(b). The values in the figures represent the mean ± standard error. In FIG. 1, the significance test was performed against the group without the extract of Aomodi leaves. * indicates p < 0.05, and *** indicates p < 0.005 (both by Dunnett's test).

[0069] As shown in FIGS. 1(a) and 1(b), the leaves of Aspergillus oryzae inhibited lipid accumulation in adipocytes in a dose-dependent manner regardless of hot water extraction or 50% ethanol extraction. The hot water extract showed a significant inhibitory effect on lipid accumulation in adipocytes at 50 μg / mL or higher, and the 50% ethanol extract showed a significant inhibitory effect at 25 μg / mL or higher.

[0070] <3-2. Evaluation of lipid accumulation in mouse body> The diet compositions are shown in Table 1. Two types of diets (control diet, Aspergillus oryzae diet) used in the experiment were prepared according to the AIN-93G composition (Table 1). As a protein source, 20% casein (milk casein, Wako Pure Chemical Industries, Ltd., Osaka) was used, as a lipid source, 7% soybean oil (Oriental Yeast Co., Ltd., Tokyo) was used, and a diet containing 30% sucrose (Dai-Nippon Meiji Sugar Co., Ltd., Tokyo) was prepared as the control diet. A diet in which 2% by weight of the control diet was replaced with the dried powder of Aspergillus oryzae leaves was used as the Aspergillus oryzae diet. At that time, the amounts of protein, lipid, dietary fiber, and carbohydrate were adjusted with casein, soybean oil, cellulose, and corn starch, respectively.

[0071] As experimental animals, 4-week-old male KK-Ay mice (CLEA Japan, Inc., Tokyo) were used. The mice were fed the control diet and after 5 days of pre-breeding, they were divided into two groups of 6 mice each so that there was no difference in body weight between the groups. The mice in each group were allowed to freely consume two types of diets (control diet, Aspergillus oryzae diet) for 5 weeks. The breeding environment was at room temperature of 22 - 23°C and a light cycle of 12 hours (8:00 - 20:00 lighting).

[0072] After the end of the breeding period, the mice were fasted for 6 hours and then, under isoflurane anesthesia, 1 mL of blood was collected from the abdominal vena cava using a syringe containing 25 μL of EDTA-2Na (1.86 mg / mL). The collected blood was centrifuged to collect the upper plasma, and erythrocyte samples were prepared from the lower layer. The plasma was stored at -80°C until analysis. The perirenal fat around the testis, mesenteric fat, and perirenal fat around the kidney were excised, and the weights of these white adipose tissues were measured.

[0073]

Table 1

[0074] The results are shown in Fig. 2. The values in the figure represent the mean ± standard error (n = 6). In Fig. 2, the significance test was performed against the group fed the control diet, and * indicates p < 0.05 (by Student's t-test). Although not shown in the figure, the body weights of the mice after 6 weeks of breeding and the measurements of ALT and AST values, which are indicators of liver function, were taken. No differences were observed among the mouse groups in each measurement, confirming that the diet containing aomori leaf powder had no effect (showed no toxicity) on the mice.

[0075] As shown in Fig. 2, the intake of aomori leaf significantly decreased the weight of intestinal mesenteric adipose tissue compared to the control (p = 0.035). The weight of peritesticular adipose tissue also showed a decreasing trend (p = 0.087) with the intake of aomori leaf.

[0076] <3-3. Evaluation of anti-inflammatory effect> The mouse macrophage-like cell line RAW264.7 cells were obtained from the RIKEN BioResource Research Center (cell number RCB0535) and cultured under the conditions of 37 °C and 5% CO2 using high-glucose D-MEM containing 10% FBS. RAW264.7 cells were seeded at 1.0×10 5 cells / cm 2They were seeded into a 96-well plate so as to obtain the desired cell density and pre-cultured for 24 hours. After 24-hour culture, the medium was changed to a phenol red-free medium, and various extracts of buckwheat leaves (Experimental Example 1 or Experimental Example 2) were mixed into the medium at final concentrations of 0, 25, 50, 100, and 200 μg / mL, followed by further culture for 24 hours. After 24-hour culture, the supernatant was removed. A phenol red-free medium was added to the control group, and 50 μL of 100 ng / mL LPS (Salmonella enterica serotype typhimurium, L6511, Merck) was added to the other groups to induce inflammation, followed by culture for 24 hours. After the culture was completed, 50 μL of the culture supernatant was transferred to a 96-well plate, and the nitric oxide (NO) concentration was measured by the Griess method. A calibration curve was prepared using NaNO2 as a standard substance. 50 μL of Griess reagent was added to 50 μL of the culture supernatant or the calibration curve preparation solution to develop color. The absorbance was measured at 550 nm using a microplate reader (Molecular Devices, LLC, CA, USA), and the NO concentration was calculated from the calibration curve.

[0077] The results are shown in Figure 3. The values in the figure represent the mean ± standard error. In Figure 3, the significance test was performed on the group treated with LPS only, and * indicates p < 0.05 (by Dunnett's test).

[0078] As shown in Figure 3, buckwheat leaves, regardless of hot water extraction or 50% ethanol extraction, suppressed NO production in macrophage cells and showed an anti-inflammatory effect in a concentration-dependent manner. In particular, for the 50% ethanol extract of buckwheat leaves, an excellent anti-inflammatory effect was confirmed at 50 μg / mL or higher.

[0079] <4. Evaluation of Sugar Absorbability> (Test 1: Evaluation of Sugar Permeability in Intestinal Epithelial Cells) The sugar permeability of the aojmodi leaf extract (50% ethanol extract) in Experimental Example 1 was measured against the small intestinal epithelial-like cell membrane. Note that <Test 1-A> in the evaluation method described below assumes ingestion of the aojmodi leaf extract before a meal, and <Test 1-B> assumes ingestion of the aojmodi leaf extract during a meal.

[0080] (Evaluation method) <Test 1-A> Caco-2 cells, a human colon cancer-derived cell line used as a small intestinal epithelial cell model, were seeded in a 24-well transwell at a seeding density of 2.5×10 5 cells and cultured in a CO2 incubator for about 3 weeks. The TEER value (trans-epithelial electrical resistance value) of the cells was measured, and after it reached 300 Ω / cm 2 500 μL of the 50% ethanol extract of the aojmodi leaf in Experimental Example 1 was added to the luminal side. The concentrations of the added aojmodi leaf extract were 100 μg / mL and 200 μg / mL. After the addition, the cells were cultured in a CO2 incubator (5% CO2, 37 °C) for 24 hours. After 24 hours, the cells were washed twice and pre-cultured in a CO2 incubator for 30 minutes. Next, a 25 mM glucose solution was added to the luminal side, and the cells were cultured in a CO2 incubator for 1 hour. After 1 hour, the solution on the basolateral membrane side was collected, and the glucose concentration was measured. The glucose concentration was measured using a Glucose Assay Kit-WST (product of Dojindo Laboratories). The obtained measurement results were calculated with the glucose concentration without the addition of the aojmodi leaf extract as the comparative example set to 100.

[0081] <Test 1-B> It was carried out in the same manner as <Test 1-A> above, except that the 50% ethanol extract of the aojmodi leaf in Experimental Example 1 adjusted to 100 μg / mL was added simultaneously when adding the 25 mM glucose solution.

[0082] (Results) The results are shown in Figs. 4 and 5. As shown in Fig. 4, the sugar permeability in <Test 1-A> where glucose was added after adding the 50% ethanol extract of Aomoji leaves in Experimental Example 1 was suppressed by 5.1% when 100 μg / mL of the Aomoji leaf extract was added, and by 10.3% when 200 μg / mL was added. Also, as shown in Fig. 5, the sugar permeability in <Test 1-B> where the 50% ethanol extract of Aomoji leaves in Experimental Example 1 was added simultaneously with glucose was suppressed by 21.9% when 100 μg / mL of the 50% ethanol extract of Aomoji leaves was added.

[0083] From the above, it was shown that adding the 50% ethanol extract of Aomoji leaves before adding glucose or simultaneously with adding glucose has an effect on suppressing sugar absorption in the small intestine.

[0084] (Test 2: Evaluation of sugar uptake in liver cells) Regarding the Aomoji leaf extracts of Experimental Example 1a (50% ethanol extract) and Experimental Example 2 (hot water extract), assuming sugar uptake in the liver, the sugar uptake amount of liver-derived cells was evaluated.

[0085] (Evaluation method) HepG2 cells, a human liver-derived cell line, were seeded in a 96-well plate at a seeding density of 9.0×10 4 cells / cm 2 and cultured in a CO2 incubator for 24 hours. Then, 1% BSA-DMEM was added and further cultured in a CO2 incubator for 24 hours. After that, 1% BSA-DMEM was removed, and the 50% ethanol extract of Aomoji leaves in Experimental Example 1a adjusted to 50 μg / mL or the hot water extract of Aomoji leaves in Experimental Example 2 adjusted to 35 μg / mL was added. After addition, it was cultured in a CO2 incubator for 24 hours and then washed. The measurement of glucose uptake was performed using the Glucose Uptake Assay Kit-Green (manufactured by Dojindo Laboratories). After adding a fluorescently labeled glucose derivative to the washed cells and culturing them in a CO2 incubator for 15 minutes, the cells were washed and the fluorescence intensity was measured at an excitation wavelength / fluorescence wavelength of 488 nm / 520 nm.

[0086] (Results) The results are shown in Figs. 6 and 7. As shown in Fig. 6, when the 50% ethanol extract of Momordica charantia leaves was added, the amount of glucose uptake increased 1.25-fold compared to the case without the Momordica charantia leaf extract. As shown in Fig. 7, when the hot water extract of Momordica charantia leaves was added, the amount of glucose uptake increased 1.14-fold compared to the case without the Momordica charantia leaf extract.

[0087] From the above, it was shown that the 50% ethanol extract of Momordica charantia leaves and the hot water extract of Momordica charantia leaves have an excellent effect on glucose uptake in the liver.

Industrial Applicability

[0088] The oral composition containing the Momordica charantia leaf extract and / or dried product of the present invention has an excellent effect on lipid accumulation suppression and anti-inflammation, and can be used by being formulated into functional foods, supplements, pharmaceutical compositions, and the like.

Claims

1. An oral composition for suppressing lipid accumulation, characterized by containing an extract and / or a dried product of Momordica charantia leaves (excluding the case where it contains Momordica charantia fruit essential oil).

2. The oral composition according to Claim 1, wherein the extract of Momordica charantia leaves is a solvent extract of Momordica charantia leaves.

3. The oral composition according to Claim 2, wherein the solvent extract of Momordica charantia leaves is a solvent extract obtained by using a mixed solvent of water and ethanol of Momordica charantia leaves as an extraction solvent.

4. The oral composition according to Claim 2, wherein the solvent extract of Momordica charantia leaves is a hot water extract of Momordica charantia leaves.

5. An oral composition for suppressing obesity, characterized by containing an extract and / or a dried product of Momordica charantia leaves (excluding the case where it contains Momordica charantia fruit essential oil).

6. An oral composition for suppressing nitric oxide production, characterized by containing an extract and / or a dried product of Momordica charantia leaves.

7. A functional food for one or more uses selected from lipid accumulation suppression, obesity suppression, and nitric oxide production suppression, containing the oral composition according to any one of Claims 1 to 6 (excluding the case where it contains Momordica charantia fruit essential oil).

8. A supplement for one or more uses selected from lipid accumulation suppression, obesity suppression, and nitric oxide production suppression, containing the oral composition according to any one of Claims 1 to 6 (excluding the case where it contains Momordica charantia fruit essential oil).

9. A pharmaceutical composition for one or more uses selected from lipid accumulation suppression, obesity suppression, and nitric oxide production suppression, containing the oral composition according to any one of Claims 1 to 6 (excluding the case where it contains Momordica charantia fruit essential oil).

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